Dr.

Bilal Al-Jaidi
Assistant Professor in Medicinal Chemistry and Drug Design
Faculty of Pharmacy, Philadelphia University-Jordan
Email: bjaidi@philadelphia.edu.jo
 Learning Outcomes
 At the end of this lesson, students will be able to:
 Define the biochemistry of Parasympathetic system.
 Classify the difference between agonist and antagonist
agents
 Outline the SAR of acetylcholine.
 Demonstrate the different chemical modification of
Acetylcholine.
 Explain the importance of Cholinergic agonists and
antagonists in disease management.
 Demonstrate the action of AchE inhibitors in the
management of Parkinson.
Overview of Nervous System
Parasympathetic Responses
Motor nerves
 Without motor nerves:

 No breathing.
 Digestion.
 Excretion.
 Muscle movement.
 Control of body temperature.
 Heart problems.
Motor nerves
1. Somatic motor NS:
 carry messages from CNS to the skeletal muscles.
 Acetylcholine is the neurotransmitter.

 Lead to muscle contraction.

Motor nerves
2. Autonomic motor NS:
 Carry messages from the CNS to smooth muscle,
cardiac muscle and adrenal medulla.
 Two subsystems:
I. 1. Parasympathetic nerves:
 From CNS to smooth muscles.
 Acetylcholine is the N.T.
II. 2. Sympathetic nerves:
 To the cardiac muscles and most of internal organs.
 Using Noradrenalin and Acetylcholine as N.T
 Adrenaline action
 increase heart rate.
 Relax GIT and UT muscles.
 Decrease salivation.
 Vasoconstriction of peripheral blood vessels.

Fight or Flight
Motor nerves
3. Enteric NS:
 Located in the wall of intestine.
 Use serotonin, neuropeptide, Acetylcholine and
Dopamine are among many other neurotransmitters
take part in this system
Nerve Impulse Transmission
Nerve Impulse Transmission
 Acetylcholine combines with cholinergic
receptors

 Nicotinic
 Excitatory response
 Muscarinic
 Excites or inhibits
 SAR for acetylcholine

Quaternary nitrogen is essential

O O

CMe3 NMe2
H3C O H3C O

Bad for activity

 SAR for acetylcholine

• Distance from quaternary nitrogen to ester is important

• Ethylene bridge must be retained

O O

H 3C O H 3C O NMe3
NMe3

Bad for activity

 SAR for acetylcholine

Ester is important

NMe3 NMe3
H 3C O H 3C

Bad for activity

 SAR for acetylcholine

Minimum of two methyl groups on quaternary nitrogen

O Et
O Et
N Me
N Et H 3C O
H 3C O Me
Et

Lower activity Active

 SAR for acetylcholine

Methyl group of acetoxy group cannot be extended

H 3C NMe3
O

Much lower activity

 SAR for acetylcholine

Conclusions:
• Tight fit between Ach and binding site
• Methyl groups fit into small hydrophobic pockets
• Ester interacting by H-bonding
• Quaternary nitrogen interacting by ionic bonding
Binding site (muscarinic)

hydrophobic
pocket Trp-307
Asp311
CH3 CH3
CO 2

N CH3
hydrophobic
O O
pockets
CH3
Trp-616 Trp-613
H
H
O N hydrophobic
pocket

Asn-617
Binding site (muscarinic)

vdw Trp-307
Asp311
CH3 CH3
CO 2
Ionic bond
N CH3 vdw
O O
H-bonds CH3 vdw
Trp-616 Trp-613
H
H
O N

Asn-617
Cholinergic Agents
 Direct acting - act on the receptors to activate a tissue
response
 Indirect acting - inhibit the action of the enzyme
Acetylcholinesterase:

 Major uses = Stimulate bladder & GI tone, constrict

pupils (miosis), neuro-muscular transmission
Hydrolysis of Acetylcholine
 Acetylcholine (Ach)

Acetyl Choline
Cholinergic receptors
1. Muscarinic receptors:
 Found in smooth muscles and cardiac muscles.
 Related to muscarine.

2. Nicotinic receptors:
 Found in skeletal muscles.
 Related to nicotine.
Subtypes of M1 M2 M3 M4 M5
M- receptor Neural Cardiac Glandular,
smooth muscles
Main CNS, Gastric & Heart , GIT, Gastric & CNS CNS
location salivary glands CNS salivary glands,
GIT, Eye
Cellular Increase IP3, Decrease in Increase IP3, As M2 As M3
response DAG, a) cAMP Ca++
Depolarization, inhibition, conductance,
excitation, b) Ca++
increase conductance,
potassium
conductance Increase K+
conductance
Functional CNS excitation, Cardiac Gastric & saliva Enhanced Not
responses gastric secretion inhibition, secretion, GI locomotion
known
neural smooth muscle
inhibition contraction,
occular
accomodation
 Cholinergic agonists

Acetylcholine as an agonist

Advantages
• Natural messenger
• Easily synthesised

Disadvantages
• Easily hydrolysed in stomach (acid catalysed hydrolysis)
• Easily hydrolysed in blood (esterases)
• No selectivity between receptor types
• No selectivity between different target organs
L-serine ethanolamine

choline Acetylcholine
Biological hydrolysis of Ach

What makes acetylcholine exceptionally prone to hydrolysis

is the possibility of folding to form an intramolecular dipole
bond that will increase the positive charge of the ester carbonyl
 Cholinergic agonists

Nicotine and muscarine as cholinergic agonists

Advantages

• More stable than Ach

• Selective for main cholinergic receptor types
• Selective for different organs

Disadvantages
• Activate receptors for other chemical messengers
• Side effects
 Cholinergic agonists

Requirements for cholinergic agonists

• Stability to stomach acids and esterases

• Selectivity for cholinergic receptors
• Selectivity between muscarinic and nicotinic receptors
• Knowledge of binding site
• SAR for acetylcholine
 Active conformation of acetylcholine

• Defines separation of ester and N

4.4A 5.9A
O O
N N

Muscarinic
Nicotinic
receptor
receptor
 Uses of cholinergic agonists

Nicotinic selective agonists

Treatment of myasthenia gravis
- lack of acetylcholine at skeletal muscle causing weakness

Muscarinic selective agonists

• Treatment of glaucoma
• Switching on GIT and urinary tract after surgery
• Treatment of certain heart defects. Decreases heart muscle
activity and decreases heart rate
 Design of cholinergic agonists

Use of steric shields

Bulky groups
Rationale

• Shields protect ester from nucleophiles and enzymes

• Shield size is important

• Must be large enough to hinder hydrolysis

• Must be small enough to fit binding site

 Acetylcholine analogues
 To overcome the instability of Ach:
 Steric shield: add large group to change the
conformation of Ach:

 3X more stable than Ach.

 More selective on muscarinic over nicotinic receptors.
 S-enantiomer is more active than the R-enantiomer
 Design of cholinergic agonists

Use of electronic factors

• Replace ester with urethane (Carbamate) group will stabilises
the carbonyl group
 Acetylcholine analogues

 Carbamate more stable ester toward hydrolysis (why?)

 NH2 and CH3 are equal sizes. Both fit the hydrophobic
pocket
 Long acting cholinergic agonist.
 Can be administered orally.
 Not selective…… just used topically in glaucoma.
 Acetylcholine analogues

 More stable.
 More selective on muscarinic receptor.
 Used to stimulate GIT and urinary bladder
after surgery.
 Muscarinic agonists

 Clinical uses:
 Treatment of glaucoma.
 Stimulate GIT and UT after surgery.
 In some heart defects.

 Pilocarpine:

 An alkaloids from Pilocarpus shrubs.

 Used in glaucoma.
 Topically only (why?)
 Muscarinic agonists
 Oxotremorine and Arecoline:

 Act on the muscarinic receptors in brain.

 Used in Alzheimer’s disease.
 Nicotinic agonists
 Mainly used in myasthenia gravis: an autoimmune
disorder in which the body produce antibodies against
cholinergic receptors.
Cholinergic Antagonists
 Of two types:
 Muscarinic antagonists:
 The prototype drug here is Atropine
Cholinergic Antagonists
 Nicotinic antagonists:
 The prototype drug is tubocurarine
MeO

Me
N
HO Me
H CH2
O

CH2
H
Me
O
H N OH

OMe
What do we mean by Antagonists
 Antagonist is a molecule that can bind to a
receptor without eliciting a response:

 it does not activate the receptor

 it will block the binding site
 Prevents the entrance of the natural ligand of
that receptor
Based on this definition
 The chemical structure of receptor antagonist does not
necessarily mimic the natural ligand structure;
Antagonist is generally bulkier,
having extra binding groups
compared to the natural ligand

Different binding mode, means no

formation of the receptor active
conformer…no response
Muscarinic Antagonists
Clinical Effects

• Decrease of saliva and gastric

secretions
• Relaxation of smooth muscle
• Decrease in motility of GIT and urinary
tract
• Dilation of pupils
Muscarinic Antagonists
Uses
• Shutting down digestion for surgery
• Ophthalmic examinations
• Relief of peptic ulcers
• Treatment of Parkinson’s Disease
• Anticholinesterase poisoning
• Motion sickness
Nicotinic Antagonists
 Normally used as muscle relaxant.

 Clinical uses:

• Neuromuscular blocker for surgical

operations
• Permits lower and safer levels of general
anaesthetic
 Muscarinic Antagonists

easily racem
Atropine

• Racemic form of hyoscyamine

• Source - roots of belladonna
• Used as a poison
• Used as a medicine
decreases GIT motility
antidote for Anticholinesterase poisoning
dilation of eye pupils
• CNS side effects – hallucinations
• Common side effects: urinary retention and blurred vision.
 Muscarinic Antagonists

• Ophthalmic use of atropine a as mydriatic (dilating) agent has been

largely replaced by use of analogs tropicamide and cyclopenatolate.

• However, atropine, and its chiral analog hyoscyamine, are utilized to

treat gastrointestinal disorders

• Also these antagonists can be used to treat the symptoms of an excess

cholinergic activity, such as the exposure to an acetylcholinesterase
inhibitor (such as a nerve gas).
 NMe3
Atropine
CH2 CH2

O CH3
C
O

• Relative positions of ester and nitrogen similar in both molecules

• Nitrogen in atropine is ionised
• Amine and ester are important binding groups (ionic + H-bonds)
• Aromatic ring of atropine is an extra binding group (vdW)
• Atropine binds with a different induced fit - no activation
• Atropine binds more strongly than acetylcholine
 Analogues of atropine

• Analogues are fully ionised

• Analogues unable to cross the blood brain barrier
• No CNS side effects
 Muscarinic Antagonists
 Hyoscine (scopolamine):

 Used in motion sickness and as antispasmodic.

 Same side effects as atropine.
 Is it more toxic than atropine?
The combination preparation ipratropium/salbutamol is a formulation
containing ipratropium bromide and salbutamol sulfate (albuterol sulfate)
used in the management of chronic obstructive pulmonary disease
(COPD) and asthma (Combivent®).
 Muscarinic Antagonists
Pharmacophore = ester + basic amine + aromatic ring

Used in excessive sweating,

spasms and nocturnal
enuresis.
 Muscarinic Antagonists
Simplified Analogues

Tropicamide Cyclopentolate Benztropine

(opthalmics) (opthalmics) O (Parkinsons disease
HN C
and ulcer)

N N
N
C O

CH2
CH
N

Benzhexol Pirenzepine
(Parkinsons disease) N
(anti-ulcer)
Me
 Tropicamide
Cyclopentolate

• Tropicamide and Cyclopentolate are among the most commonly employed

mydriatic (dilating) and cycloplegic (paralyzing) agents

• Both function as antagonists at the muscarinic acetylcholine receptors

 Muscarinic Antagonists
SAR for Antagonists

Important features
• Tertiary amine (ionised) or a quaternary nitrogen
• Aromatic ring
• Ester
• N-Alkyl groups (R) can be larger than methyl (unlike agonists)
• Large branched acyl group
• R’ = aromatic or heteroaromatic ring
• Branching of aromatic/heteroaromatic rings is important
 Muscarinic Antagonists
SAR for Antagonists

Me Me
CH O
Me
O CH2CH2 N CH C
O C Me
Me CH2 O CH2 CH2NR2
O
Cl

Active Inactive
Proposed Binding Site for Antagonists

C O

O
H2N Asn
CH2

CH2

NMeR2
CO 2
 Nicotinic Antagonists
Curare

• Extract from curare plant (Chondodendron tomentosum)

• Used for poison arrows
• Causes paralysis (blocks acetylcholine signals to muscles)
• Active principle = tubocurarine
MeO

Me
N
HO Me
H CH2
O

CH2
H
Me
O Tubocurarine
H N OH

OMe
Native Indians hunting animals with this poison
were able to eat the animal's contaminated flesh
without being affected by the toxin because
tubocurarine cannot easily cross mucous
membranes and is thus inactive orally.
 Nicotinic Antagonists
Pharmacophore

• Two quaternary centres at specific separation (1.15nm)

• Different mechanism of action from atropine based
antagonists
• Different binding interactions

Clinical uses

• Neuromuscular blocker for surgical operations

• Permits lower and safer levels of general anaesthetic
• Tubocurarine used as neuromuscular blocker
 Nicotinic Antagonists
Analogues of tubocurarine

• Long lasting
• Long recovery times
• Side effects on heart
• No longer in clinical use
• Esters incorporated
• Shorter lifetime (5 min)
• Fast onset and short duration
• Frequently used during intubation
• Side effects at autonomic ganglia
 Nicotinic Antagonists
Analogues of tubocurarine

• Steroid acts as a spacer for the quaternary centres (1.09nm)

• Acyl groups are added to introduce the Ach skeleton
• Faster onset than tubocurarine but slower than suxamethonium
• Longer duration of action than suxamethonium (45 min)
• Pancuronium is used to block the neuromuscular junction during surgery or
intubation.

• In the US, Pancuronium (Pavulon) is the second of three drugs used in execution by
lethal injection
 Nicotinic Antagonists
Analogues of tubocurarine

• Design based on tubocurarine and suxamethonium

• Lacks cardiac side effects
• Rapidly broken down in blood both chemically and metabolically
• Avoids patient variation in metabolic enzymes
• Lifetime is 30 minutes (Self destruct system limits lifetime)
• Administered as an i.v. drip
 Atracurium instability

• Atracurium is stable under acid pH

• But unstable under blood pH due to
Hofmann elimination
Analogues of tubocurarine

• Faster onset (2 min)

• Shorter duration (15 min)
No chance for Hoffmann elimination (WHY?)
 Anticholinesterase agents
 Lead to Ach accumulation… have cholinergic effect.
Types of cholinesterase enzymes
 Acetylcholinesterase
Located in synapses
Substrate selectivity:
 Ach
 Plasma cholinesterase
Located in plasma (non-neuronal)
Substrate selectivity:
 Ach
 Succinylcholine
 Local anesthetics (procaine)
Anticholinesterase agents
R Relative Potency
CH3 1.0
C2H5 5.0
C3H7 100
C4H9 50

 Tetraalkylammonium ion
 Bind to anionic site and block Ach binding
 Reversible
Anticholinesterase agents
 Anticholinesterase agents
 Either reversible or irreversible inhibition.
1. Carbamate:

 Systemically toxic (why?).

 Use in Glaucoma, myasthenia gravis and Alzheimer’s.
 Used as antidote for atropine poisoning (why?)
 SAR:
 Carbamate is essential.
 Benzene ring is important.
 Pyrrolidine ring is important.
 What are the possible side effects?
 Anticholinesterase agents
 Physostigmine analogues:

 has CNS side effects.

 Neostigmine has less CNS side effects and more stable

than miotine (why).
 Both used in myasthenia gravis.
 Anticholinesterase agents

 Same profile as neostigmine.

 was used by troops to protect against nerve gases.
 Is it orally available?
 In which dosage form will be administered?
Irreversible Acetylcholinesterase inhibitors

 Organophosphates
 Irreversible binding to Cholinesterase active site
 Longer acting
 Used in the treatment of glaucoma
Irreversible Acetylcholinesterase inhibitors

 Organophosphates Nerve gases

 Irreversible binding to AchE
Irreversible Acetylcholinesterase inhibitors

 Organophosphates Insecticides
 Irreversible binding to AChE
 Rapidly inactivated in mammals compared to insects
Carboxyesterase
Mammals, Birds
Antidote for AchE “poisoning”

 Pralidoxime chloride
(Protopam;PAM)
 Antidote for pesticide or
nerve gas poisoning
 Most effective if given
Cl- within a few hours of
exposure
Clinical Uses of acetylcholinesterase
inhibitors
1. An introduction to Medicinal Chemistry by Graham L. Patrick.
4th edition, Oxford, 2009
2. Wilson and Gisvolds text book of organic medicinal and
pharmaceutical chemistry by John H. Black and John M. Beale,
jr. 12th edition, Lippincott Williams and Wilkings 2011.
3. Foyes principle of medicinal chemistry by David H. Williams,
Thomas L. Leuke, Williams O. Foye. Lippincott William and
Wilkins. 7th edition, 2013.
The End